http://bura.brunel.ac.uk/handle/2438/32871 2026-04-18T03:46:24Z 2026-04-18T03:46:24Z Mensah, NE Efthymiou, A Mureanu, N Martín Monreal, MT Vaikkinen, H Kannambath, S Bowman, A Menon, A Tree, T Lombardi, G Dhami, P Nicolaides, KH Scottà, C Shangaris, P http://bura.brunel.ac.uk/handle/2438/33116 2026-04-09T02:00:59Z 2026-04-03T00:00:00Z

Title: Single-cell transcriptomics identifies regulatory T cell heterogeneity in gestational diabetes mellitus Authors: Mensah, NE; Efthymiou, A; Mureanu, N; Martín Monreal, MT; Vaikkinen, H; Kannambath, S; Bowman, A; Menon, A; Tree, T; Lombardi, G; Dhami, P; Nicolaides, KH; Scottà, C; Shangaris, P Abstract: Background: Gestational diabetes mellitus (GDM) is a common pregnancy complication associated with hyperglycaemia, chronic inflammation and adverse health outcomes. Regulatory T cells (Tregs) are thought to contribute to GDM due to their role in suppressing inflammation. However, whether specific Treg subsets are transcriptionally dysregulated in patients with GDM remains unclear. Methods: To investigate Treg transcriptional variation in GDM, we applied single-cell RNA sequencing to Tregs and CD4 + T cells isolated from the blood of 13 healthy pregnant women and 10 female patients with GDM. Results: We observed no significant differences in Treg cluster proportions with disease status, however, Memory CD4 + T cells were more abundant in patients diagnosed with GDM, substantiated by mass cytometry. We report Treg subsets altered in GDM, including naive Tregs with reduced expression of AP-1 transcription factor subunits and effector Tregs with increased signalling of genes associated with angiogenesis. Expression levels of genes dysregulated in GDM Tregs were informative of GDM status in pseudobulk, placental and whole blood mRNA from independent cohorts. TXNIP, which regulates glucose levels, emerged as the most significant discriminator of GDM status from bulk mRNA. Conclusions: This study uncovers transcriptional differences of Treg cell subsets from GDM patients and transcriptional markers informative of GDM status. Description: Plain Language Summary: Gestational diabetes mellitus (GDM) is a common pregnancy condition linked to high blood sugar and increased inflammation, which can affect the health of both mother and baby. Immune cells called regulatory T cells (Tregs) help control inflammation, and their activity in a mother’s blood may be linked to GDM. To understand how Tregs behave in patients with GDM, we captured these cells from blood samples of pregnant women diagnosed with GDM and pregnant women without a GDM diagnosis. We profiled the expression of RNA in individual Tregs from these patients. We found that, while overall Treg numbers are similar, the activity of specific genes varies in Tregs from women with GDM. Disrupted RNA levels of one gene related to glucose control (TXNIP) may be an informative marker for GDM in blood. Our findings enhance the understanding of immune changes in GDM and may inform future approaches for early detection and monitoring.; Data availability: Single-cell RNA sequencing data has been submitted to Gene Expression Omnibus (Accession: GSE280975). Bulk RNA sequencing data was downloaded from Gene Expression Omnibus (Accession: GSE154414; GSE92772 - RNA sequencing data of whole blood cells of normal glucose tolerant (NGT) and gestational diabetes (GDM) pregnant women). Data underlying the figures are available on Zenodo [59.] Mensah, N. Single-cell transcriptomics identifies regulatory T cell heterogeneity in Gestational Diabetes Mellitus [Data set]. Zenodo. https://doi.org/10.5281/zenodo.18032075 (2026).; Code availability: R scripts used to perform the analyses are available at GitHub [60.] Mensah, N. E. rutepo_gdm_treg. GitHub repository. https://github.com/NMNS93/rutepo_gdm_treg (2025).; Springer Nature is providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.; Supplementary information is available online at: https://www.nature.com/articles/s43856-026-01563-0#Sec24 .

2026-04-03T00:00:00Z Edwards, W Sun, N Wang, Y Lu, Y Wang, C Mastronicola, D Scottà, C Romano, M Cejas, CM Espinet, A Lombardi, G Chiappini, C http://bura.brunel.ac.uk/handle/2438/32944 2026-04-07T11:08:03Z 2026-03-05T00:00:00Z

Title: Perfusion microbioreactor for CAR-Treg manufacturing Authors: Edwards, W; Sun, N; Wang, Y; Lu, Y; Wang, C; Mastronicola, D; Scottà, C; Romano, M; Cejas, CM; Espinet, A; Lombardi, G; Chiappini, C Abstract: Summary: Manufacturing cell and gene therapies (CGTs) at scale presents challenges in cost, product consistency, and adaptability to personalised treatments. Traditional large-volume bioreactors are designed to support cell growth through controlled nutrient delivery and gas exchange, but are poorly suited to the decentralised, small-batch production required for personalised therapies like Chimeric Antigen Receptor (CAR) T-cells. To address this, we have developed the KCL-Microbioreactor (K-MBR), a closed microbioreactor platform based on microfluidic principles. Engineered in polydimethylsiloxane (PDMS), the K-MBR combines spatial confinement, semi-continuous perfusion, and integrated viral transduction in a compact footprint enabling efficient gene delivery and robust expansion of therapeutic cells. We demonstrate the platform’s utility by generating functional CAR-Tregs targeting HLA-A2, achieving a 92% increase in yield compared to conventional methods. The K-MBR offers a streamlined, solution for CGT manufacturing, with potential to reduce productions cost and enhance scalability across a broad range of cell therapies. Description: Highlights: • Perfusion microbioreactor achieves Treg expansion comparable to gold standard G-Rex device. • Spatial confinement increases lentiviral transduction efficiency of primary, human cells. • Compact, low-volume platform reduces the physical footprint of cell manufacturing. • Device supports future automation and advances progress toward point-of-care production.; Data and code availability: All data reported in this paper will be shared by Ciro Chiappini upon request; this paper does not report original code. All datasets generated and analysed in this study, including raw flow cytometry files and source data, are available from the lead contact upon reasonable request.; Supplemental Information is available online.

2026-03-05T00:00:00Z Busharat, Zabreen http://bura.brunel.ac.uk/handle/2438/32769 2026-02-24T13:17:24Z 2025-01-01T00:00:00Z

Title: Novel memory phenotype Tfh cells arise without overt antigen stimulation and are important for adaptive immune responses against viral infection Authors: Busharat, Zabreen Abstract: Pathogen-induced memory Tfh cells exert a Tfh effector response during reinfection, regulating the generation of high-affinity antibodies. Here, we define novel memory-phenotype Tfh cells which are generated from naïve T cells under homeostatic conditions. These MP Tfh cells are phenotypically and functionally similar to pathogen-induced Tfh cells. MP Tfh cells can be defined by Tfh cell specific markers, CXCR5, BCL6, and PD-1, and markers of pathogen-induced long lived Tfh cells, FR4. T-bethigh MP T cells exert an innate-like Th1 response against viral infections. The transcription factor EGR2 is a repressor of T-bet function, and we found that MP Tfh cells are distinct from T-bethigh MP T cells but express EGR2 highly. Previously, we found Egr2 is required for MP T cell homeostasis and inflammation. Here, we observed that, in Egr2/3-/- CD4+ MP T cells, MP Tfh cell development is impaired. FR4+ EGR2 + MP T cells upregulate genes related to homeostatic proliferation, Tfh cell development and metabolic pathways of pathogen-induced memory Tfh cells. MP Tfh cells can exert an adaptive function by regulating B cell-mediated IgG production in vitro whereas MP Tfr cells are involved in suppressing MP Tfh cell function, thereby preventing excessive inflammation. In vivo, MP Tfh cells support germinal centre formation and induce neutralising antibody production after infection with vaccinia virus. Thus, MP Tfh cells with similar characteristics to pathogen-induced memory Tfh cells are developed in absence of environmental antigens and to date are the only CD4+ MP T cell subset associated with an adaptive immune response against viral infection. Description: This thesis was submitted for the award of Doctor of Philosophy and was awarded by Brunel University London

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